Microarray dispensing with real-time verification and inspection

ABSTRACT

A microarrayer for spotting solution onto a receiving surface in an automated microarray dispensing device. Elements of the present invention include: at least one dispense head for spotting the receiving surface, at least one light source capable of illuminating the receiving surface, at least one camera operating in conjunction with the at least one light source. The at least one camera is capable of acquiring and transmitting surface image data to a computer. The computer is programmed to receive the surface image data and analyze it. The computer will then generate post analysis data based on the analysis of the surface image data. The post analysis data is available for improving the spotting of the solution onto the receiving surface. In a preferred embodiment, the surface image data includes information relating to receiving surface alignment, information relating to spot quality, and receiving surface identification information. In a preferred embodiment, the analysis of the information relating to receiving surface alignment enables the computer to make automatic adjustments to the relative positions of the at least one dispense head and the receiving surface to increase the accuracy of the spotting. In a preferred embodiment, the analysis of the information relating to spot quality identifies a spot as pass or fail. An operator is then able to rework the spot. In a preferred embodiment, the analysis of the receiving surface identification information enables the computer to track each receiving surface. In a preferred embodiment the receiving surface is a plurality of slides.

The present invention relates to automated microarray dispensingdevices, more specifically it relates to automated microarray dispensingdevices with automated quality inspection capability. This applicationis a continuation in part application of U.S. patent application Ser.No. 09/611,256 filed Jul. 6, 2000, now U.S. Pat. No. 6,558,623, which isincorporated by reference herein.

BACKGROUND OF THE INVENTION

Microarrays, also known as biochips, have recently become important inthe study of genomics. The use of a microarray involves laying down anordered array of genetic elements onto a solid substrate such as aslide. Depending on the application, a microarray may consist of genomicDNA, reverse-transcribed cDNA, or smaller chains of oligonucleotides aswell as any preparatory substrates. The microarray is useful because itallows genetic analysis to take place on a massively parallel scale,wherein thousands of genes and markers can be scored in one experiment.

A microarrayer, also known as a DNA array printer, is a high-capacitysystem used to print a microarray onto slides. Typically, a microarrayeris a specially built robotic platform designed to transfer solutionsfrom the well of some type of microplate onto another surface foranalysis. This process of depositing the liquid spot onto the slide isknown as “spotting”.

Recently, microarrayers have become extremely popular in laboratoriesbecause they add to the efficient productivity of the laboratory to beable to print samples onto slides accurately and rapidly. Affymetrix,Inc., with offices in Santa Clara, California, makes an automatedarrayer called the 417 ARRAYER (Part No. 0-0001 and Part No. 0-0009).BioRobotics, with offices in Boston, Massachusetts, produces twoversions of an automated arrayer called the MICROGRID and MICROGRID II.GeneMachines, with offices in Menlo Park, Calif. makes an arrayer calledthe OMNIGRID (Model No. OGR-02). Packard Instrument Company with officesin Meriden, Conn. makes an automated arrayer called the BIOCHIP ARRAYER.

Although there are some differences between each of the above listedmicroarrayers, they are all similar in that they each spot microarraysin an automated fashion. However, there are significant problems withthe prior art devices that detracts from their efficient operation.

A first problem arises due to the fact that as blank slides are cycledthrough prior art microarrayers, they can become askew or positionedimproperly underneath dispensing tips. This problem results in spotsbeing positioned improperly on the slides. A second problem can ariseeven if the slide is positioned correctly under the dispensing tips. Itis possible for the spot to be deposited in the correct position, but beof poor quality and therefore useless as far as experimentationpurposes.

Up to now, the only way to deal with these problems was to have a humanoperator visually monitor and inspect the microarrayer during itsoperation or inspect the samples after they come off the machine. Thissolution is an unacceptable waste of human effort. The BIOCHIP ARRAYERmade by Packard Instrument Company has attempted to deal with theproblem of monitoring the spotting process. However, it has only limitedverification functionality with its integrated camera. This means thatit verifies whether or not a spot has been dispensed, without anyquality inspection to analyze whether that spot was good or bad.

What is needed is a better microarrayer with automated qualityinspection capability.

SUMMARY OF THE INVENTION

The present invention provides a microarrayer for spotting solution ontoa receiving surface in an automated microarray dispensing device.Elements of the present invention include: at least one dispense headfor spotting the receiving surface, at least one light source capable ofilluminating the receiving surface, at least one camera operating inconjunction with the at least one light source. The at least one camerais capable of acquiring and transmitting surface image data to acomputer. The computer is programmed to receive the surface image dataand analyze it. The computer will then generate post analysis data basedon the analysis of the surface image data. The post analysis data isavailable for improving the spotting of the solution onto the receivingsurface. In a preferred embodiment, the surface image data includesinformation relating to receiving surface alignment, informationrelating to spot quality, and receiving surface identificationinformation. In a preferred embodiment, the analysis of the informationrelating to receiving surface alignment enables the computer to makeautomatic adjustments to the relative positions of the at least onedispense head and the receiving surface to increase the accuracy of thespotting. In a preferred embodiment, the analysis of the informationrelating to spot quality identifies a spot as pass or fail. An operatoris then able to rework the spot. In a preferred embodiment, the analysisof the receiving surface identification information enables the computerto track each receiving surface. In a preferred embodiment the receivingsurface is a plurality of slides.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the major components of a preferred embodiment of thepresent invention.

FIG. 2 shows slides fifty slides located on 5 locating plates after theslides have been spotted.

FIGS. 3–32 illustrate the sequence of operation of a preferredembodiment of the present invention.

FIG. 33 shows the major components of a preferred embodiment of thepresent invention.

FIG. 34A shows the rework dispense head in the up position.

FIG. 34B shows the rework dispense head in the down position.

FIG. 34C shows a slide with a 2D bar code.

FIGS. 35A and 35B shows dispense tips attached to dispense heads.

FIG. 36 shows a preferred embodiment of the present invention mounted ona vibration isolated base.

FIG. 37 shows the major components of a preferred embodiment of thepresent invention.

FIGS. 38A and 38B show a flowchart for the programming of a preferredembodiment of the present invention.

FIGS. 39A and 39B show the reworking of a slide for a preferredembodiment of the present invention.

FIG. 40 shows another preferred embodiment of the present invention.

FIG. 41 shows another preferred embodiment of the present invention.

FIG. 42 shows another preferred embodiment of the present invention.

FIG. 43 shows another preferred embodiment of the present invention.

FIG. 44 shows a dispense head over a microplate.

FIGS. 45A–45C show a pre-spot slide.

FIG. 46 shows a dispense head over a slide.

FIG. 47 shows a preferred incubator.

FIG. 48 shows robotic plate handling.

FIGS. 49A–49B show a microplate.

FIGS. 49C–49D show a dispense head over a microplate.

FIGS. 50 and 51 show a preferred microplate and dispense head.

FIG. 52A shows another preferred embodiment of the present invention.

FIG. 52B shows microplates in storage racks.

FIG. 53 shows spots deposited on a preferred slide.

FIGS. 54–58 show a preferred monitor screen display.

FIG. 59 shows spot inspection data.

FIG. 60 shows dispense pin spot inspection analysis.

FIG. 61 shows a calibration target.

FIG. 62 shows another preferred monitor screen display.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

A detailed description of a preferred embodiment of the presentinvention can be described by reference to FIGS. 1–38B. During theoperation of the present invention, solution from reservoir plate 5 isautomatically deposited onto an array of fifty blank slides 4A1–4E10located on locating plates 4A–4E (see FIG. 1). An operator is able toselect via a computer interface whether dispense tip 42 (locatedunderneath dispense head 40) or a 4×6 array of dispense tips 7 (locatedunderneath dispense head 6) will be used to make the deposits ontoslides 4A1–4E10. In the preferred embodiment, dispense tips 7 and 42 arequill type dispense tips. Locating plates 4A–4E are mounted on linearactuator 15 so that they can move along the x-axis. Linear actuator 26is mounted on linear actuator 21 so that linear actuator 26 can movealong the y-axis. Dispense heads 6 and 40 are mounted on linear actuator26 so that they can move along the z-axis. Camera 12 with strobe light13 is focused so as to permit recording of the deposition process andfunctions to permit verification of slide identification information andto permit verification of proper deposition of solution on the slides.Periodically, during the cycle, the dispense tips are cleaned in soniccleaner 9, rinsed in rinse fountain 10, and then dried in vacuummanifold 11. After the solution has been deposited onto the slides, theoperator can retrieve locating plates 4A–4E containing slides 4A1–4E10from the position shown in FIG. 2.

Sequence of Operation of a Preferred Embodiment

FIGS. 3–32 illustrate the sequence of operation of a preferredembodiment of the present invention.

In a preferred embodiment of the present invention, the operation of thecomponents is controlled by PC control system 300, as shown in FIG. 37.FIGS. 38A–38E show a flowchart representing preferred programming of PCcontrol system 300 and corresponds to the sequence illustrated in FIGS.3–32.

As shown in FIG. 3, an operator places five locating plates 4A–4E eachhaving ten clean, blank slides 4A1–4E10 on platform 2. In a preferredembodiment of the present invention, slides 4A1–4E10 are made bySequonem with offices in San Diego, Calif. A preferred slide is shown inFIG. 16B. It has ninety-six etched dispense positions 60 and has its ownunique 2D bar code 65 for identification purposes.

As shown in FIG. 4, linear actuator 15 moves platform 2 so that slide4A1 is underneath the dispense head 6. Dispense head 6 is positioneddirectly above slide 4A1. Using camera 12 and the strobe light 13, animage is acquired of slide 4A1. The camera reads the bar code andinspects the positioning and alignment of slide 4A1 on locating plate3A. The software then analyzes the position data and stores theinformation. The information stored and will be used later to adjust thepositions of slide 4A1 and dispense head 6 to ensure accurate placementof the solution on the slide.

After camera 12 has acquired the image of slide 4A1, linear actuator 26is moved via linear actuator 21 to the position shown in FIG. 5 so thatdispense head 6 is directly above slide 4A2. Using camera 12 and thestrobe light 13, an image is acquired of slide 4A2. As with slide 4A1,camera reads the bar code and inspects the positioning and alignment ofslide 4A2 on locating plate 4A. The software then analyzes the positiondata and stores the information. The information stored and will be usedlater to adjust the positions of slide 4A2 and dispense head 6 to ensureaccurate placement of the solution on the slide.

Linear actuator 26 is then moved via linear actuator 21 to the positionshown in FIG. 6 so that dispense head 6 is directly above sonic cleaner9.

As shown in FIG. 7, mounting plate 25 moves downward via linear actuator26 so that dispense tips 7 are dipped in sonic cleaner 9 for aprogrammable time period while the cleaner is turned on. When finished,mounting plate 25 moves upward as shown in FIG. 8.

Then, as shown in FIG. 9, platform 31 moves to the left via pneumaticslide 30, thereby moving rinse fountain 10 and vacuum manifold 11 to theleft. Linear actuator 26 is moved via linear actuator 21 back so that itis directly above rinse fountain 10.

As shown in FIG. 10, mounting plate 25 moves downward via linearactuator 26 so that tips 7 are dipped in rinse fountain 10 for aprogrammable time period while the fountain is turned on.

After rinsing, mounting plate 25 moves upward via linear actuator 26, asshown in FIG. 11. Platform 31 moves to the right via pneumatic slide 30,thereby moving rinse fountain 10 and vacuum manifold 1 to the right.

As shown in FIG. 12, dispense tips 7 are then lowered into the vacuummanifold 11 via linear actuator 26 and the vacuum is turned on for aprogrammable time period, thereby drying dispense tips 7. This cleaningcycle can be set by the user, via the computer interface, to be repeatedas many times as necessary.

Mounting plate 25 is then raised via linear actuator 26 as shown in FIG.13. Linear actuator 26 is then moved via linear actuator 21 so thatdispense head 6 is directly above reservoir plate 5, as shown in FIG.14.

Mounting plate 25 is then lowered via linear actuator 26 so that tips 7are dipped into the solution contained in reservoir plate 5, as shown inFIG. 15. While in reservoir plate 5, dispense tips 7 pick up some of thesolution to be dispensed.

Linear actuator 26 then moves to the first dispense position shown inFIG. 16A so that dispense head 6 is above slide 4A1. Based on theearlier positioning data regarding slide 4A1 (see discussion of FIG. 4),linear actuator 21 makes minute positioning adjustments to linearactuator 26 and linear actuator 15 makes minute positioning adjustmentsto platform 2 in order to accurately position dispense head 6 over slide4A1 at the first dispense position.

FIG. 16B shows a top view of a blank slide 4A1. In this preferredembodiment, slide 4A1 has 96 positions 60 arranged in an 8×12 array. Ateach position 60, slide 4A1 is etched so as to be better able to retaina drop of solution deposited at the spot.

Dispense head 6 is then lowered via linear actuator 26 so that tips 7are in contact with slide 4A1 at the first dispense position, as shownin FIG. 17A. As tips 7 contact slide 4A1, spots 62 are placed on slide4A1 via surface tension, as shown in FIG. 17B.

Dispense head 6 is raised via linear actuator 26, as shown in FIG. 18.Based on the earlier positioning data regarding slide 4A1, linearactuator 21 makes minute positioning adjustments to linear actuator 26and linear actuator 15 makes minute positioning adjustments to platform2 in order to accurately position dispense head 6 over slide 4A1 at thesecond dispense position.

Dispense head 6 is then lowered via linear actuator 26 so that tips 7are in contact with slide 4A1 at the second dispense position, as shownin FIG. 19A. As tips 7 contact slide 4A1, more spots 62 are added toslide 4A1, as shown in FIG. 19B.

Dispense head 6 is raised via linear actuator 26, as shown in FIG. 20.Based on the earlier positioning data regarding slide 4A1, linearactuator 21 makes minute positioning adjustments to linear actuator 26and linear actuator 15 makes minute positioning adjustments to platform2 in order to accurately position dispense head 6 over slide 4A1 at thethird dispense position.

Dispense head 6 is then lowered via linear actuator 26 so that tips 7are in contact with slide 4A1 at the third dispense position, as shownin FIG. 21A. As tips 7 contact slide 4A1, more liquid spots 62, areadded to slide 4A1, as shown in FIG. 21B.

Dispense head 6 is raised via linear actuator 26, as shown in FIG. 22.Based on the earlier positioning data regarding slide 4A1, linearactuator 21 makes minute positioning adjustments to linear actuator 26and linear actuator 15 makes minute positioning adjustments to platform2 in order to accurately position dispense head 6 over slide 4A1 at thefourth dispense position.

Dispense head 6 is then lowered via linear actuator 26 so that tips 7are in contact with slide 4A1 at the fourth dispense position, as shownin FIG. 23A. As tips 7 contact slide 4A1, more liquid spots 62 are addedto slide 4A1, as shown in FIG. 23B.

Dispense head 6 is raised via linear actuator 26, as shown in FIG. 24.Camera 12 and strobe 13 scans slide 4A1 and acquires images and inspectsfor spot quality. It is at this point that PC control system 300 (FIG.37) identifies slide 4A1 as pass or fail. (Preferred computer controlledtechniques for making this determination are discussed in a followingsection.)

As shown in FIG. 25, based on the earlier positioning data regardingslide 4A2 (see discussion regarding FIG. 5), linear actuator 21 makespositioning adjustments to linear actuator 26 and linear actuator 15makes positioning adjustments to platform 2 in order to accuratelyposition dispense head 6 over slide 4A2 at the first dispense positionfor slide 4A2.

The four-stage liquid dispense cycle (explained above with respect toslide 4A1 in discussion regarding FIGS. 17A–24) is repeated so that atthe end of the four-stage cycle, slide 4A2 contains spots 62, as shownin FIG. 26B. At the end of the four stage cycle, dispense head 6 israised via linear actuator 26, as shown in FIG. 26A. Camera 12 andstrobe 13 scans slide 4A2 and acquires images and inspects for spotquality. It is at this point that the control system identifies slide4A2 as pass or fail.

As shown in FIG. 27, linear actuator 21 moves linear actuator 26 so thatdispense head 6 is above slide 4A3.

The sequence outlined in the discussion regarding slides 4A1 and 4A2(depicted in FIGS. 4–26 ) is repeated with regards to slides 4A3 and4A4. To summarize, by utilizing light provided by strobe 13, camera 12will first record the positions of slides 4A3 and 4A4. Then, dispensetips 7 are dipped in sonic cleaner 9. Dispense tips 7 are then rinsed inrinse fountain 10. Then, dispense tips 7 are dried in vacuum manifold11. This cycle is repeated as needed. Then, liquid is picked up bydispense tips 7 when dispense tips 7 are lowered into reservoir plate 5.Then, liquid is spotted onto slide 4A3 by dispense tips 7 in afour-stage liquid dispense cycle. Likewise, liquid is spotted onto slide4A4 in a four-stage liquid dispense cycle so that liquid has beenspotted on both slides, as shown in FIGS. 28B and 28C.

The process is then repeated for the remaining six slides 4A5–4A10 untilall the slides on locator plate 4A have been spotted, as shown in FIG.29.

Linear actuator 15 then moves platform 2 so that slide 4B1 of locatorplate 4B underneath dispense head 6, as shown in FIG. 30.

In a similar fashion, dispense tips 7 continue to spot all ten slides onlocator plates 4B–4E, until the last slide 4E10 has been spotted, a

After slide 4E10 has been spotted and camera 12 and strobe 13 hasscanned slide 4E10 for spot quality, linear actuator 15 moves platform 2to the position shown in FIG. 32 so an operator can remove locatorplates 4A–4E.

Computer Controlled Pass-Fail Determination Technique

The computer controlled pass-fail determination technique determinesindividual spots as pass or fail based on several criteria. For eachslide, the camera system scans a region to look for a spot. In apreferred embodiment the criteria that are applied to that inspectionregion are spot presence, spot size in area, spot location, and spotgeometry. Additional criteria can be added through softwareconfiguration. Each of the criteria can have upper and lower limitsdesignated which define the acceptable values for that particularcriteria. Any value that falls outside of the limits for any criteriaqualifies that spot and slide as failed. The actual inspection valuesare determined by analyzing the grayscale intensity of each pixel. Thetotal number of pixels falling above and below a threshold are talliedto give values for each of the inspection criteria.

Rework Capability

As explained above, after each slide has been spotted, camera 12 andstrobe 13 scans the slide and acquires images and inspects for spotquality. It is at this point that the control system identifies theslide as pass or fail. In a preferred embodiment of the presentinvention, an operator monitoring the spotting process via monitor 305(FIG. 37) has the option of correcting a slide that has failed.

For example, FIG. 29 shows locating plate 4A after all slides 4A1–4A10have been spotted. At this point, an operator can scan locating plate4A. A good plate shows up green as in all slides pass. A plate with atleast one bad spot on one of the slides shows up red. The user can thenzoom in on the bad slide and the good and bad spots show up green andred respectively as pass or fail. From there, the user can decidewhether or not to rework the bad spots.

FIGS. 39A and 39B show where an operator has decided to rework slide 4A6that has a spot that has failed quality inspection. Dispense head 40 islowered via pneumatic slide 41 so that dispense tip 42 is lower thandispense tips 7. Solution from reservoir plate 5 is then deposited onthe slide at the location of the failed spot. If there are other spotsthat failed, the operator can likewise rework those spots in a similarfashion.

Although in the description given above regarding the rework process,the operator reworked failed slides after locating plate 4A had beenentirely spotted, it is also possible to rework failed slides at otherstages during the spotting process. For example, it may be desirable towait until all slides 4A1–4E10 on plates 4A–4E have been spotted (FIG.31) before reworking them. This allows an operator to be free to doother activities while the initial spotting is taking place. Then, afterall slides have been spotted, he can come back and do all the reworkingat one sitting.

Alternatively, it may be desirable to rework each slide immediatelyafter it has been spotted, as shown in FIG. 24.

Automatic Rework Capability

The previous section described a preferred embodiment where an operatorcan decide whether or not to rework a spot based on a computerdetermination of pass or fail. In another preferred embodiment therework decision is made automatically by the computer based on whetheror not the spot has passed or failed. In this preferred embodiment, thecomputer makes a determination whether or not a spot has passed orfailed using the computer controlled pass-fail determination techniqueearlier described. If, based on its analysis, the computer determinesthat the spot has failed, the computer will automatically take steps torework the spot. For example, dispense tip 42 will extract solution fromreservoir plate 5. Then, the computer will lower dispense head 40 viapneumatic slide 41 so that dispense tip 42 is lower than dispense tips7, as shown in FIGS. 39A and 39B. Solution from reservoir plate 5 willthen deposited on the slide at the location of the failed spot.

Components of a Preferred Embodiment of the Present Invention Three AxisRobotic Positioning Stage

In a preferred embodiment, linear actuators 26, 21 and 15 are industrialgrade precision ground ball screw linear actuators, as shown in FIG. 33.These linear actuators are manufactured by Parker Automation (Model#s:404XR and 406XR series). They are each controlled by a smartservomotor (SmartMotor, Model #2320 VRE, manufactured by Animatics, withoffices in Santa Clara Calif.), which is a fully self-contained closedloop servo system. Each of these smart servomotors contains the motor,encoder, amplifier, and controller all in one small package mounted tothe linear actuator. Linear actuator 15 (the x-axis positioning device)has an overall travel distance of 600 mm with an accuracy within+/−0.032 micrometers Linear actuator 21 (the y-axis positioning device)has an overall travel distance of 400 mm with an accuracy within+/−0.032 micrometers. Linear actuator 26 (the z-axis positioning device)has an overall travel distance of 100 mm with an accuracy within +/−micrometers. In this preferred embodiment, this extreme accuracy isneeded to accommodate very small spot size and spacing between spots.The linear actuators have pitches of 5mm per revolution giving apositioning accuracy of 0.032×10⁻⁶ meters.

Linear actuator 15 controls the positioning of platform 2 containingslides 4A1–4E10 along the x-axis of motion making all slides presentableto the dispense head 6. Linear actuator 21 controls the positioning ofthe dispense head along the y-axis of motion making all slides 4A1–4E10,sonic cleaner 9, rinse fountain 10, vacuum manifold 11, and reservoirplate 5 presentable to dispense head 6. Linear actuator 26 controls thepositioning of dispense head 6 along the z-axis of motion allowingdispense head 6 to be lowered to and raised from all slides 4A1–4E10,sonic cleaner 9, rinse fountain 10, vacuum manifold 11, and reservoirplate 5.

Cleaning Station

Cleaning station 33 consists of sonic cleaner 9, rinsing fountain 10,and a drying vacuum manifold 11. In a preferred embodiment, soniccleaner 9 is an ultrasonic cleaner manufactured by Prosonic, Inc. (partno. E0028). Sonic cleaner 9 can contain either a cleaning solution orsimply purified water. Dispense tips 7 are dipped in the sonic cleaner9, where the ultra sonic oscillations of the cleaning solution clean thetips.

Rinsing fountain 10 and the vacuum manifold 11 are placed on a pneumaticslide 30. Pneumatic slide 30 is used to select which operation is to beperformed, rinsing or drying. The reason for this slide is so that bothoperations can be performed at a single position along the y-axis. Thisallows for both operations without having to increase the overall travelof linear actuator 26 along the y-axis.

Rinsing fountain 10 pumps in purified water and drains it out to a wastebin. Dispense tips 7 are dipped in this purified water to rinse away anydebris or cleaning solution that may remain on the tips after cleaning.

Drying vacuum manifold 11 is a block with an array of holes in it thatmatch the array of dispense tips 7. The tips are inserted into theblock, each of these holes are connected to a manifold which isconnected to a vacuum generator and air supply. The vacuum pulls awayany remaining liquid or debris left on the dispense tips after rinsing.

Dispense Head Assemblies

As shown in FIG. 35B, dispense head 6 is a 4×6 grid Micro Quill Holder(part no. 11946-0) made by Major Precision of Arizona. A 4×6 array ofprimary dispense tips 7 are held in dispense head 6. As shown in FIG.35A, dispense head 40 is a Micro Quill Holder also made by MajorPrecision. Dispense tip 42 is held in dispense head 40. Dispense tips 7and 42 are spring loaded within the dispense heads 6 and 40.

As shown in FIGS. 1, 34A and 34B, dispense head 6 is rigidly mounted tomounting plate 25, whereas dispense head 40 is mounted to pneumaticslide 41 (manufactured by Robohand, Inc., with offices in Montroe,Conn., part no. MPS1-2). Mounting plate 25 is capable of moving up anddown along the z-axis via linear actuator 26. Furthermore, dispense head40 is capable of independent additional movement up and down along thez-axis via pneumatic slide 41, as shown in FIGS. 34A–34B.

During normal operation, such as that depicted in the sequenceillustrated in FIGS. 3–32, dispense head 6 is used to spot slides4A1–4E10. F front view of dispense tips 7 in contact with slide 4A1.Dispense tips 7 will be used to spot slide 4A1 at positions 60, as shownin FIG. 34C. Note that when dispense head 6 has been selected, dispensehead 40 is raised via pneumatic slide 41 so that dispense tips 42 do notinterfere with the spotting process.

If, however, the operator wishes to spot slide 4A1 at positions 61 (FIG.34C), dispense head 40 will be lowered via pneumatic slide 41 so thatdispense tips 42 are in contact with slide 4A1 and dispense tips 7 areout of the way, as shown in FIG. 34B.

Camera and Lighting

In a preferred embodiment, camera 12 and strobe 13 are mounted to theside of linear actuator 26 as shown in FIG. 1. Camera 12 is aself-contained camera with image processing and Ethernet capabilitiesmanufactured by DVT Corporation with offices in Norcross, Ga. (series600 model). Using light provided by strobe 13, camera 12 can snappictures while in dynamic motion, process the image for results, passthe results off to the PC control system, and prepare for the next imageacquisition. The camera uses a 55 mm Telecentric lens which provides theproper field of view and magnification for reading of 2D bar code 62(FIG. 34C) and for image inspection. Strobe light 13 is preferably ModelDL2449, manufactured by Advanced Illumination, with offices inRochester, Vt. In the preferred embodiment, the image acquisition timeis ˜40 ms and the image processing time is ˜50 ms. The system can alsobe equipped with a flouresence device along with the camera for furthergenomic expression analysis.

Vibration Isolated Base

As shown in FIG. 36, vibration isolated base 80 is provided to minimizeany possible affects that high frequency environmental vibrations mighthave on the dispensing process. This base is a pneumatic system whichacts as a shock absorber to the system.

In a preferred embodiment, the base is manufactured by Newport, Inc.with offices in Irvine, CA, model # CM-225.

PC Based Control System

FIG. 37 depicts a block diagram of PC control system 300 and othercomponents of a preferred embodiment of the present invention. PCControl System 300 includes CPU 301 with associated memory (RAM 302 andROM 303). It also includes a touch screen monitor/interface 305 thatallows for operator monitoring, intervention and control of the presentinvention. In the preferred embodiment, the computer system is a PCbased computer equipped with an ethernet card and running windowssoftware. The programming is preferably written in VISUAL BASIC. (VISUALBASIC is a federally registered trademark of Microsoft Corp., a DelawareCorporation) PC Control System 300 is equipped with CMS (CentralMonitoring System). The CMS gives PC Control System 300 it's own IP(Internet Protocol) address and ethernet connectivity. This allows forremote monitoring and control via Intranets as well as Internet providedthat the bandwith is available for proper functionality. The software ishighly comfigurable to allow increased flexibility for customers withvarying slide types, slide sizes, slide orientations, spot size, spotspacing and many other variables.

Control of the Components of the Preferred Embodiment of the PresentInvention through the PC Control System

As previously stated, linear actuators 26, 21 and 15 are industrialgrade precision ground ball screw linear actuators. As shown in FIG. 37,PC control system 300 sends signals to smart servomotors 26A, 21A and15A to control linear actuators 26, 21 and 15, respectively. PC controlsystem 300 controls sonic cleaner 9 and rinse fountain 10. Compressedair source 310 provides compressed air to pneumatic slides 30 and 41 viavalves 310 controlled by PC control system 300. Vacuum generator 320provides a vacuum to vacuum manifold 11 via valve 317. As previouslyexplained camera 12 and strobe 13 work in conjunction to provide sensorydata to PC control system 300. This input is used to accurately positionthe dispense heads over the slides to ensure optimum spotting and toverify the quality of the spotting as “pass” or “fail” using multiplecriteria as to placement at intended location as well as spot size (toobig or too small).

Second Preferred Embodiment of the Present Invention

A second preferred embodiment of the present invention is shown in FIG.41. In the second preferred embodiment, dispense head 106 is connectedvia mounting plate 125 to linear actuator 126 so that dispense tips 107can be raised and lowered along the z-axis into solution in microplate190. Camera 112 and strobe 113 are rigidly mounted to the side of linearactuator 126 so that they remain stationary with respect to the side oflinear actuator 126 along the z-axis. Linear actuator 126 is mounted tolinear actuator 121 so that it can move along the y-axis.

Platform 182 is mounted to linear actuator 180 so that it can move alongthe x-axis. Locating plate 4A is placed on top of platform 182. Platform102 is mounted to linear actuator 115 so that it can move along thex-axis. Microplate 190 is place on top of platform 102. In thispreferred embodiment platform 102 has the capacity to hold tenmicroplates 190.

Solution in microplate 190 is removed via dispense tips 107. Linearactuator 126 then moves along the y-axis so that dispense tips 107 areabove locating plate 4A. The solution is then spotted in a fashionsimilar to that described for the earlier preferred embodiments. Camera112 with strobe 113 is focused so as to permit recording of thedeposition process and functions to permit verification of slideidentification information, permit verification of proper deposition ofsolution on the slides, and to verify slide alignment. As explainedabove, slide image data is transferred via camera 112 to a PC controlsystem where the data is analyzed. The results of the analysis are thenavailable for improving the spotting of the solution onto the slides.For example, spots that have failed to meet the threshold limits can bereworked. Also, the computer can automatically make adjustments to therelative positions of the slides and dispense tips based on the slidealignment analysis. Periodically, during the cycle, the dispense tipsare cleaned in sonic cleaner 109, then rinsed in the rinse fountain anddried in vacuum manifold 111. Operation of the First PreferredEmbodiment with the Second Preferred Embodiment

The first preferred embodiment (described in the sequence illustrated inFIGS. 3–32) can be used in conjunction with the second preferredembodiment to spot slides. For example as shown in FIG. 32, locatingplate 4A can be removed from the microarrayer via an operator after ithas been spotted with a base solution. Locating plate 4A can betransferred to the microarrayer depicted in FIG. 41. It can be placed onplatform 180. DNA from microplate 190 can then be spotted on top of thebase solution spotted already on slides 4A1–4A10.

Use of the Present Invention with Other Microarrayers

Although the present invention was described as being used with thepreferred microarrayer depicted in the sequence described by referenceto FIGS. 3–32, those of ordinary skill in the art will recognize that itis possible to use camera 12, strobe 13 and a PC control system inconjunction with a variety of microarrayer designs. For example, in thebackground section of this application, several microarrayers werementioned. It would be possible to one of ordinary skill in the art tomodify a prior art automatic microarrayer to include camera 12 andstrobe 13. Camera 12 and strobe 13 would then work in conjunction toprovide sensory data to PC control system 300, as described above. Also,as explained above, the input would be used to accurately position thedispense heads over the slides to ensure optimum spotting and to verifythe quality of the spotting as “pass” or “fail”.

Modification of Rework Dispense Tips

The previous embodiments showed one dispense tip 42 extending downwardfrom dispense head 40. It was explained how the single dispense tip 42is used for reworking (correcting) defective spots. It is possible,however, to modify dispense head 40 so that multiple dispense tips canextend downward from dispense head 40. A preferred embodiment is shownin FIG. 40 in which five dispense tips 42A–E extend down below dispensehead 40. In this preferred embodiment, dispense tips 42A–E areretractably connected to dispense head 40. As shown in FIG. 40, dispensetips 42A–D are retracted inside dispense head 40. The rightmost dispensetip 42E is extended below the other dispense tips and is spotting slide4A1. In a preferred embodiment, dispense tips 42A–E are mounted to apneumatic slides 43.

An advantage of this embodiment is that each dispense tip 42 can beconfigured to dispense a different volume of solution. For example, in apreferred embodiment, dispense tip 42A would dispense 1 nL of solution,dispense tip 42B would dispense 2 nL of solution, dispense tip 42C woulddispense 4 nL of solution, dispense tip 42D would dispense 8 nL ofsolution, and dispense tip 42E would dispense 16 nL of solution.

After initially spotting the slides as explained above, camera 12 andstrobe 13 would work in conjunction to provide sensory data to PCcontrol system 300 reporting the quality of the spots. The spots wouldthen be classified as pass or fail. If a spot has failed, the softwarein conjunction with PC control system 300 would determine the amount ofsolution required to correct the failed spot. Then, during the reworkingsequence, the dispense tip that dispenses the most correct volume wouldbe extended down from dispense head 40 and the other dispense tips wouldbe retracted upward inside dispense head 40, as shown in FIG. 40.

Third Preferred Embodiment

A third preferred embodiment of the present invention is seen byreference to FIG. 42. FIG. 42 shows a perspective view of microarrayer250. In the third preferred embodiment, dispense head 202 is connectedvia mounting plate 213 to linear actuator 214 so that dispense tips 208can be raised and lowered along the z-axis. Camera 215 and strobe 216are also mounted to mounting plate 213 and can be raised and loweredalong the z-axis by linear actuator 214. Linear actuator 214 is mountedto linear actuator 217 so that it can move along the y-axis.

Platform 218 is mounted to linear actuator 219 so that it can move alongthe x-axis. 140 slides 220(1)–220 (140) are mounted on top of platform218. Microplates 204(A–C) and pre-spot slide holder 205 having pre-spotslide 206 are placed on top of platform 218.

Operation of the Third Preferred Embodiment Removing Solution From theMicroplate

To remove solution from microplate 204(A), linear actuator 219 movesplatform 218 along the x-axis and linear actuator 217 moves linearactuator 214 along the y-axis so that dispense head 202 is positionedover microplate 204(A), as shown in FIG. 44. Linear actuator 214 thenlowers dispense head 202 so that dispense tips 208 are dipped into thewells of microplate 204(A). Linear actuator then raises dispense head202. Dispense tips 208 contain solution removed from microplate 204(A).

Pre-Spotting the Solution onto a Pre-Spot Slide

The third preferred embodiment includes pre-spot slide 206. Pre-spotslide 206 allows excess solution on the dispense tips to be removedprior to the solution being spotted onto slides 220(1)–220 (140). Thisis achieved by repeatedly pressing the dispense tips onto the pre-spotslide until the excess solution is removed.

To pre-spot solution onto pre-spot slide 206, linear actuator 217 moveslinear actuator 214 along the y-axis so that dispense head 202 ispositioned above pre-spot slide holder 205 having pre-spot slide 206, asshown in FIGS. 45A and 45B. Linear actuator 214 then lowers mountingplate 213. As previously stated, dispense head 202 is mounted tomounting plate 213. Linear actuator lowers mounting plate 213 withdispense head 202 until dispense tips 208 come into contact with thesurface of pre-spot slide 206, as shown in FIG. 45C. The act of dispensetips 208 pressing against the surface of pre-spot slide 206 causesexcess solution on the sides of the dispense tips to come off thedispense tips. The third preferred embodiment is programmed so thatlinear actuator 214 raises and lowers dispense head 202 in a repetitivefashion so that dispense tips 208 are pressed against pre-spot slide 206repetitively until the desired amount of solution is removed. The amountof times dispense tips 208 should be pressed varies depending on theproperties of the solution being deposited and can be modified by theuser by inputting instructions into computer 280 (FIG. 42).

Spotting Solution onto Slides

After the solution has been pre-spotted onto pre-spot slide 206, thesolution can then spotted onto selected microplates 220(1)–220 (140) ina fashion similar to that described above in reference to the first andsecond preferred embodiments. For example, to spot solution onto slide220(1), linear actuator 219 moves platform 218 along the x-axis andlinear actuator 217 moves linear actuator 214 along the y-axis so thatdispense head 202 is positioned over slide 220(1), as shown in FIG. 46.The solution is then spotted in a fashion similar to that described forthe earlier preferred embodiments. Camera 215 with strobe 216 is focusedso as to permit recording of the deposition process and functions topermit verification of slide identification information, permitverification of proper deposition of solution on the slides, and toverify slide alignment. As explained above, slide image data istransferred via camera 215 to a PC control system where the data isanalyzed. The results of the analysis are then available for improvingthe spotting of the solution onto the slides. For example, spots thathave failed to meet the threshold limits can be reworked. Also, thecomputer can automatically make adjustments to the relative positions ofthe slides and dispense tips based on the slide alignment analysis.Periodically, during the cycle, the dispense tips are cleaned in soniccleaner 231, then rinsed in the rinse fountain 230 and dried in vacuummanifold 232.

Robotic Loading of Microplates

FIG. 43 shows robotic plate loader 252 positioned between microarrayers250. Microarrayers 250 and robotic plate loader 252 are controlled byprogrammable computer 280.

As shown in FIG. 43, microplate staging area 254 has 4 microplatestorage racks 255(A)–255(D). Each storage rack is capable of holding upto 15 microplates. Robotic plate loader 252 is capable of removing themicroplates from each of the storage racks. Robotic plate loader 252 canpivot so that arm 257 can be extended towards a desired storage rack.Robotic gripper 256 can then grip the desired plate and remove it fromthe storage rack. The microplate can then be placed on either of theplatforms 218(A)–(B) (as shown) or at pre-staging area 258. Microplatelids 259 can then be removed and placed at lid holding areas 260 and261.

Robotic Plate Loading from Incubator

By storing microplates in an incubator, the temperature of the solutioninside the microplates can be kept at a controlled level. FIGS. 47 and48 show another preferred embodiment in which microplates stored inincubator 270 are automatically loaded via robotic arm 271 onto platform218. FIG. 48 shows a top view and FIG. 47 shows a perspective view ofincubator 270.

In FIG. 48, incubator access door 278 has been raised permitting outsideaccess to incubator 270. Fifteen microplates are stored inside storageracks 274(A)–274(D). Robotic pivot arm 275 of robot 272 inside incubator270 is capable of pivoting about axis 273 so that it can reach storageracks 274(A)–274(D).

FIG. 48 shows robotic pivot arm 275 in dotted line positioned so thatgripper 276 is in front of storage rack 274(A). Robotic arm 275 is shownin solid line in a vertical position placing microplate 274(A)(13) onaccess plate 277.

In FIG. 47, robotic arm 271 has maneuvered gripper 279 so that it wasable to remove microplate 274(A)(13) from access plate 277, rotate it 90degrees, and place it on platform 218 as shown.

Automatic Spotting from Multiple Microplates onto Multiple Slides

In the third preferred embodiment, microarrayer 250 can be programmed toremove solution from multiple microplates and spot the solution ontomultiple slides 220 (1–140). FIG. 52A shows a top view of platform 218.Microplates 204(A), 204(B) and 204(C) are on platform 218. Microplates204(D–O) are in storage rack 255(A). Microplates 285(A–O) are in storagerack 255(B). Microplates 286(A–O) are in storage rack 255(C) andmicroplates 287(A–O) are in storage rack 255(D), as shown in FIG. 52AB.

Robotic plate loader 252 is positioned between platform 218 and storageracks 255(A–C) and is capable of accessing the positions occupied bymicroplates 204(A-C) and any of the microplate storage positions instorage racks 255(A–D). Dispense head 202 (see also FIG. 42) is capableof being positioned over microplates 204(A–C) via microarrayer 250 andis capable of removing solution from microplates 204(A–C) and spottingthat solution on any of the slides 220(1–140). Robotic plate loader 252can move microplates 204(A–C) and store them back in storage rack 255(A)and then place other microplates onto platform 218. For example, amicroarrayer 250 could be programmed to place microplates 204(A–C) backinto storage rack 255(A) and then place microplates 285(A), 286(D), and287(H) onto platform 218.

Flexible Mapping

In the preferred embodiment, a user of microarrayer 250 (FIG. 42) isable to customize the manner in which spots are deposited on slides220(1–140).

FIG. 49A shows a top view of 384 well microplate 204A. In the preferredembodiment, microplate 204A is divided into 8 sections 204A1–204A8, asshown in FIG. 49B. Each section 204A1–204A8 covers a 4×12 array ofwells. In one preferred embodiment, a unique solution is deposited ineach section 204A1–204A8.

FIG. 50 shows a top view of 4×12 pin dispense head 202 next tomicroplate 204(A). Dispense tips P1–P48 are shown as “+” signs and arearranged as shown. The 48 dispense tips on dispense head 202 are spacedto fit into the wells of each 4×12 well section A1–A8 of microplate 204A(FIGS. 51 and 49B). FIG. 49C shows dispense head 202 positioned oversection 204A2 and FIG. 49D shows dispense head 202 positioned oversection 204A5.

FIG. 53 shows a top view of slide 220(1). Dispense head 202 havingdispense tips P1–P48 dispenses the spots onto slide 220(1). Eachdispense tip P1–P48 is responsible for dispensing the spots in its own8×8 array. For example, dispense tip PI dispenses all the spots in array283(1) and dispense tip P4 dispenses all the spots in array 283(4),(FIG. 53).

In the preferred embodiment, a user can program computer 280 so thatdispense head 202 will remove solution from section A1–A8 of microplate204A and deposit the solution in a customized fashion on slide 220(1).

For example, in a preferred embodiment a user will use computer 280 tocontrol microarrayer 250 and will see on monitor 281 a screen imagesimilar to that depicted in FIG. 54. At the upper left corner of thescreen the user inputs the number of microplates for spotting. In theexample shown, the user has indicated eight plates. Eight microplateswill provide solution to fill one slide with 3072 spots. For example, asshown in FIG. 53 slide 220(1) holds forty-eight 8×8 arrays of spots.Each 8×8 array has 64 spots. Since there are 48 arrays on slide 220(1),there are a total of 48×64=3072 spots on slide 220(1).

Each microplate has eight 4×12 sections from which dispense head 202 canremove solution. Since there are eight microplates from which dispensehead 202 can remove solution and each microplate has eight sections,there are a total of 8×8=64 sections from which solution can be removed.The total number of sections is displayed in the upper left corner ofthe screen in FIG. 54.

Customized Mapping of the Arrays

In mapping the forty-eight 8×8 arrays to be deposited on slide 220(1) itis only necessary for the user to focus on how one of the forty-eightarrays will appear. This is because each array 283(1–48) is identical tothe other. For example, as shown in FIG. 53, while dispense tip P1 ofdispense head 202 (FIG. 50) is depositing solution to position Al ofarray 283(1), dispense tip P2 and P28 are depositing solution from thesame microplate section to positions A1 of array 283(2) and A1 of array283(28), respectively. Moreover, while dispense tip P1 is depositingsolution to position C6 of array 283(1), dispense tip P32 is depositingsolution from the same microplate section to position C6 of array283(32). Therefore, after all 64 spots have been deposited by P1 ontoarray 283(1), dispense tips P2–P48 have simultaneously deposited 64spots to each array 283(2–48) for a total of 3072 spots.

To map an 8×8 array, the user selects from which plates he wants toremove solution by entering the plate numbers into boxes 300, as shownin FIG. 54. In the present example the user wants to remove solutionfrom each section of microplates 204(A), 204(B), 204(C), 204(D), 285(C),286(D), 286(H), and 287(B). To select which microplate he wants to firstmap, the user mouse clicks on one of the eight microplates at the bottomof the screen.

In FIG. 55, the user has selected microplate 204(A) to map. Microplate204(A) appears in the upper center of the screen under the heading“Current Plate”. The user then mouse clicks on a section that he wantsto map. In the present example the user has mouse clicked on section204A1 under the “Current Plate” heading. This causes the 204A1 sectionto darken. The user then mouse clicks on position A1 of spot array map301. The numerals 204A1 then appear in the Al position of the array toindicate that the A1 position has been mapped and section 204A1 ofmicroplate 204(A) at the left bottom corner of the screen has darkenedto indicate that the 204A1 section has been mapped.

In FIG. 56, the user has mouse clicked on section 204A2 under the“Current Plate” heading. This causes the 204A2 section to darken. Thenthe user mouse clicks on C4 of spot array map 301. The numerals 204A2then appear in the C4 position of the array to indicate that the C4position has been mapped and section 204A2 of microplate 204(A) at theleft bottom corner of the screen has darkened to indicate that the 204A2section has been mapped.

In a similar fashion, the user can map the rest of microplate 204(A)onto spot array map 301. FIG. 57 shows microplate 204(A) completelymapped onto spot array map 301.

The user can continue to map the rest of the microplates by followingthe above-described procedures with respect to each of the seven othermicroplates. For example, FIG. 58 shows spot map array 301 completelymapped.

After having filled in spot map array 301, the user saves his customizedmapping and runs microarrayer 250 (FIG. 42). Microarrayer 250 willremove solution from microplates 204(A–C) first because they arepositioned on platform 218 so that they can be accessed by dispense head202. After solution has been removed from each of the eight sections ofa microplate, robotic plate loader 252 (FIG. 52A) will return themicroplate to its appropriate storage rack. The robotic plate loaderwill then place the next microplate at the position vacated by theprevious microplate on platform 218. For example, after solution hasbeen removed from microplate 204(A), robotic plate loader 252 willreturn microplate 204(A) to its slot in storage rack 255(A) (FIGS. 52Aand 52B). Robotic plate loader 52 will then place microplate 204(D) atthe spot vacated by microplate 204(A). Microarrayer will continue tospot slide 220(1) until it has been completed spotted, as shown in FIG.53.

If the user would like to spot more slides with the pattern shown inspot map array 301 in FIG. 58, he inputs into computer 280 the number ofslides he would like to spot. For example, if he would like to spot allof the slides 220(1–140) the user inputs that information into thecomputer and microarrayer 250 will spot slides 220(1–140). Each slidewill be spotted in the same pattern as the other slide. For example,each slide will be spotted in accordance with the pattern shown in spotmap array 301 in FIG. 58. Similarly, if the user just wants to spotslides 220(1–10), he inputs that information into computer 280 andslides 220(1–10) will be spotted in accordance with the pattern shown inspot map array 301 in FIG. 58.

Spot Inspection Data and Spot Inspection Statistics

As stated previously, camera 215 with strobe 216 is focused so as topermit recording of the deposition process and spot quality andfunctions to permit verification of slide identification information,permit verification of proper deposition of solution on the slides, andto verify slide alignment. Spot data covering the qualities of each spoton the slides is transferred via camera 215 to a computer 280 systemwhere the data is analyzed and stored. In a preferred embodiment, thisinformation is displayed in table format to the user via monitor 281.

FIG. 59 shows a preferred spot inspection data table. Information suchas spot size and spot offset is reported. Status indicating the spot aseither good or bad is reported as well as whether rework was requiredfor the spot.

FIG. 60 shows a preferred table reporting spot inspection statistics. Inthis table, the user can quickly ascertain the quality of dispense tipsP1–P48 on dispense head 202. For example, by referring to FIG. 60, theuser can see that dispense tip P1 deposited 20 spots and they were allgood spots and rework was not required. In contrast, the user can seethat dispense tip P2 also deposited 20 spots but that only 15 of themwere good. Rework was required for 5 spots. The user can then determinethat dispense tip P1 is operating effectively, but that dispense tip P2might need to be cleaned, repaired or replaced. Preferably, the tableshown in FIG. 60 also reports information such as spot diameterstatistics for each dispense tip and spot offset statistics for eachdispense tip.

Exportation of Mapping Information, Spot Inspection Data and SpotInspection Statistics

In a preferred embodiment, it is possible to export to another computerthe data associated with the spotting process. For example, as discussedabove under the headings “Flexible Mapping” and “Customized Mapping ofthe Arrays” the user can customize the mapping process in accordancewith his wishes. The mapping information can then be saved by computer280 and matched with the appropriate slide for later reference. Forexample, if slide 220(1) was mapped in accordance with spot map array301 shown in FIG. 58, then an individual later working with slide 220(1)could refer to the saved mapping information and immediately know thesource microplates for each spot on the slide. Likewise, it is alsopossible to save and export the spot inspection data and spot inspectionstatistics discussed above for later reference.

In one preferred embodiment, the saved information is exported via acomputer network to another computer on the computer network. In anotherpreferred embodiment, the saved information is saved to a computer discand the computer disc is physically transferred to another computerwhere it can be accessed.

Calibration Target

Preferably, microarrayer 250 (FIG. 42) includes calibration target 303(see also FIGS. 52 and 61). As shown in FIG. 61, calibration target 303includes array 304. Array 304 has a known dimension. Also, the size ofthe dots of the array is known as well as the distance between the dots.The color of calibration target 303 is also known. To calibrate camera215, the user positions it over calibration target 303. Camera 215 isfocused onto array 304 and measurements are taken. The measurements arecompared to the known value. If there are discrepancies, thenadjustments can be made to camera 215 to calibrate it. For example, ifthere is a discrepancy between the known values and the size of array204, or the size of the dots, or the spacing of the dots, then zoom lensof the camera can be adjusted. Or, if there is a discrepancy between theknown color of the calibration target and the reported color (i.e., toolight or too dark), the iris or the aperture of the camera can beadjusted.

Extra Sonic Cleaning and Vacuum Stations

In the preferred embodiment, microarrayer 250 includes two vacuumstations 232 and two sonic cleaning stations 231 (FIG. 52A). By havingtwo sets of stations, the user is better assured of thoroughly cleaningdispense tips P1–P48. Preferably, one sonic station 231 contains waterand the other sonic station 231 contains alcohol.

Parameter Adjustments for Spotting

In a preferred embodiment, the user is able to adjust parametersrelating to spot quality via computer 280 (FIG. 42). On monitor 281, theuser will see a screen image similar to that shown in FIG. 62. The usercan adjust load parameters and dispense parameters by entering valuesinto boxes 305. The definitions of the adjustable parameters are givenbelow in Table 1:

TABLE 1 Load Dwell The amount of time the dispense tips are immersed inthe well solution before lifting up. Load Offset The distance (inmicrons) from the bottom of the wells that the dispense tips stop whileimmersed in the well solution. (Ex: −500 microns means that the dispensetips stop 500 microns above the bottom of the well.) Load Speed Thespeed of the dispense tips as they move up and down over the microplatewells. Load Repeat The number of times the loading of the dispense tipsis repeated as they are positioned over the wells. Load Repeat Theamount of time in the up position before the loading Delay of thedispense tips is repeated. Dispense/Load The number of times thedispense tips dispense solution before they are loaded again. DispenseThe amount of time the dispense tips are in contact with Dwell the slidebefore lifting up. Dispense The distance (in microns) the springs of thedispense tips Offset compress before lifting up. Dispense The speed ofthe dispense tips as they move up and down Speed over the slide.Dispense The number of times the dispensing of solution Repeat (i.e.,spotting) is repeated as the dispense tips are positioned over theslide. Disp Repeat The amount of time in the up position before theDelay dispensing of solution is repeated.

While the above description contains many specifications, the readershould not construe these as limitations on the scope of the invention,but merely as exemplifications of preferred embodiments thereof. Thoseskilled in the art will envision many other possible variations arewithin its scope. For example, although the above sequence described adispensing process utilizing slides that have 96 dispense positions,those of ordinary skill in the art will recognize that it is possible touse other slides as well. For example, 384 or 1536 position slides couldbe used. It is also possible to use a blank microscope slide with nopre-etched dispense positions. Accordingly, the location of thedifferent dispense positions will vary depending on the type of slidebeing used. The spacing and orientation of the slide can be selected byan operator through the maintenance menu on the computer interface.Also, the previous embodiments disclosed using a strobe light toilluminate the slide below the camera. One of ordinary skill in the artwill recognize that it is possible to illuminate the slide with otherlight sources besides a strobe light. For example, the slide could beilluminated with a camera flash, a constant bright light, or afluorescence device, such as a fluorescent LED. Additionally, the slidecould be illuminated from the side opposite the camera. If afluorescence device is used to illuminate the slide, those of ordinaryskill in the art will recognize that it is possible to add a fluorescentdye to the solution being spotted to achieve more in depthcharacterizations. For example, by using a fluorescent LED and addingfluorescent dye to the solution, greater volume determination can beachieved based on fluorescent intensity of the spot. Also, in thepreferred embodiment, it was mentioned that dispense tips 7 and 42 werequill type dispense tips, it would be obvious to substitute other typesof dispense tips. For example, piezo type dispense tips could also beused. The present invention was taught showing the microarrayerdepositing solution onto a plurality of slides. One of ordinary skill inthe art would recognize that the microarrayer could be used to depositsolution onto receiving surfaces that include a variety of slide typesand surfaces other than slides. For example, the microarrayer coulddeposit solution onto a glass slide, or a slide whose surface has beentreated to enhance the spotting or the experimental results, or slidesthat have porous or semi-porous surfaces, membrane slides, gold coatedslides, slides made of porous silicon, or any other surface that may bebeneficially printed upon. Additionally, while the present invention wastaught using substantially flat slides, it would be obvious to usesurfaces that are slightly to moderately curved to print upon.Accordingly the reader is requested to determine the scope of theinvention by the appended claims and their legal equivalents, and not bythe examples which have been given.

1. A microarrayer for spotting solution onto a receiving surface,comprising: A. a receiving surface, B. at least one dispense head forspotting said receiving surface, C. at least one light source capable ofilluminating said receiving surface, D. at least one camera operating inconjunction with said at least one light source, said at least onecamera capable of acquiring and transmitting surface image data, E. acomputer programmed to: 1) receive said surface image data from said atleast one camera, 2) analyze said surface image data, and 3) generatepost analysis data based on said analysis of said surface image data,wherein said post analysis data comprises information relating to thesuccess or failure of said microarrayer to successfully spot solutiononto said receiving surface, F. an adjustment device for permittingadjustments to be made to said spotting of solution onto said receivingsurface, wherein said adjustments are based on said post analysis data.2. The microarrayer as in claim 1, wherein said receiving surface is aplurality of receiving surfaces.
 3. The microarrayer as in claim 1,wherein said receiving surface is a plurality of slides.
 4. Themicroarrayer as in claim 1, wherein said adjustment device is areworking device for permitting the microarrayer operator to rework aspot via the microarrayer based on said post analysis data.
 5. Themicroarrayer as in claim 1, wherein said adjustment device is areworking device for permitting said computer to rework a spot via themicroarrayer based on said post analysis data.
 6. The microarrayer as inclaim 1, wherein said microarrayer further comprises: A. at least onedispense tip for immersion into said solution, B. a pre-spot receivingsurface, wherein said microarrayer removes excessive amounts of saidsolution by pressing said at least one dispense tip against saidpre-spot receiving surface.
 7. The microarrayer as in claim 1, whereinsaid microarrayer removes said solution from a plurality of microplates,wherein a robot automatically positions said microplates withinaccessible reach of said microarrayer.
 8. The microarrayer as in claim7, wherein said plurality of microplates are stored in storage racks ata storage location, wherein said robot is capable of retrieving saidmicroplates from said storage racks and returning said microplates tosaid storage racks.
 9. The microarrayer as in claim 8, wherein saidstorage location is an incubator.
 10. The microarrayer as in claim 1,further comprising a flexible mapping device for allowing themicroarrayer operator to customize the pattern in which said dispensehead deposits solution onto said receiving surface.
 11. The microarrayeras in claim 1, wherein said post analysis further comprises spotinspection data comprising information about the characteristics ofindividual spots.
 12. The microarrayer as in claim 1, wherein saiddispense head comprises a plurality of dispense tips, wherein said postanalysis data further comprises spot inspection statistics comprisinginformation about the performance of said dispense tips.
 13. Themicroarrayer as in claim 1, wherein said microarrayer further comprisesa calibration target for calibration of said camera.
 14. Themicroarrayer as in claim 1, wherein said microarrayer further comprises:A. at least two vacuum stations, and B. at least two sonic cleaningstations.
 15. The microarrayer as in claim 1, further comprising: A. aplurality of dispense tips, and B. a second adjustment device forallowing the microarrayer operator to adjust the manner in whichsolution is loaded onto said plurality of dispense tips and to adjustthe manner in which solution is deposited onto said receiving surface.16. A microarrayer for spotting solution onto a receiving surface,comprising: A. a receiving surface means, B. a dispensing means forspotting said receiving surface means, C. a light source means capableof illuminating said receiving surface means, D. a camera meansoperating in conjunction with said light source means, said camera meanscapable of acquiring and transmitting surface image data, E. a computermeans programmed to: 1) receive said surface image data from said atleast one camera means, 2) analyze said surface image data, and 3)generate post analysis data based on said analysis of said surface imagedata, wherein said post analysis data comprises information relating tothe success or failure of said microarrayer to successfully spotsolution onto said receiving surface means. F. an adjustment means forpermitting adjustments to be made to said spotting of solution onto saidreceiving surface means, wherein said adjustments are based on said postanalysis data.
 17. The microarrayer as in claim 16, wherein saidreceiving surface is a plurality of receiving surfaces.
 18. Themicroarrayer as in claim 16, wherein said receiving surface is aplurality of slides.
 19. The microarrayer as in claim 16, wherein saidadjustment means is a reworking means for permitting the microarrayeroperator to rework a spot via the microarrayer based on said postanalysis data.
 20. The microarrayer as in claim 16, wherein saidadjustment means is a reworking means for permitting said computer torework a spot via the microarrayer based on said post analysis data. 21.The microarrayer as in claim 16, wherein said microarrayer furthercomprises: A. at least one dispense tip for immersion into saidsolution, B. a pre-spot receiving surface means, wherein saidmicroarrayer removes excessive amounts of said solution by pressing saidat least one dispense tip against said pre-spot receiving surface means.22. The microarrayer as in claim 16, wherein said microarrayer removessaid solution from a plurality of microplates, wherein a robot meansautomatically positions said microplates within accessible reach of saidmicroarrayer.
 23. The microarrayer as in claim 22, wherein saidplurality of microplates are stored in storage racks at a storagelocation, wherein said robot means is capable of retrieving saidmicroplates from said storage racks and returning said microplates tosaid storage racks.
 24. The microarrayer as in claim 23, wherein saidstorage location is an incubator.
 25. The microarrayer as in claim 16,further comprising a flexible mapping means for allowing themicroarrayer operator to customize the pattern in which said dispensingmeans deposits solution onto said receiving surface.
 26. Themicroarrayer as in claim 16, wherein said post analysis date furthercomprises spot inspection data comprising information about thecharacteristics of individual spots.
 27. The microarrayer as in claim16, wherein said means comprises a plurality of dispense tips, whereinsaid post analysis data further comprises spot inspection statisticscomprising information about the performance of said dispense tips. 28.The microarrayer as in claim 16, wherein said microarrayer furthercomprises a calibration target means for calibration of said camera. 29.The microarrayer as in claim 16, wherein said microarrayer furthercomprises: A. at least two vacuum stations, and B. at least two soniccleaning stations.
 30. The microarrayer as in claim 16, furthercomprising: A. a plurality of dispense tips, and B. a second adjustmentmeans for allowing the microarrayer operator to adjust the manner inwhich solution is loaded onto said plurality of dispense tips and toadjust the manner in which solution is deposited onto said receivingsurface means.